Airway epithelial cells suppress antigen-specific T cell proliferation.

<p>A: Gating strategy for assessment of T cell proliferation by CFSE dilution. All CD4+ cells having a lower CFSE intensity than non-proliferated control cells were designated as “CFSE-low” cells. Proliferation of OVA-specific DO11.10 T cells was induced by DC-mediated OVA-presentation or T cell-depleted splenocyte ( = APC) presentation of pOVA for the indicated time (B) or on day 4 (C–E). Cultures w/o antigen served as controls. Epithelial-mediated suppression was assessed in co-culture with the LA4-cell line, a murine, type II pneumocyte cell line. B: Pulmonary epithelial cells inhibit antigen-induced T cell proliferation. C: Epithelial-mediated suppression of T cell proliferation is independent from type of antigen-presenting cell. Epithelial-mediated suppression of T cell proliferation is almost entirely dependent on cell-contact of epithelial cells with T cells and DC (D) or APC (E). TC  =  T cell, DC  =  dendritic cell, OVA  =  Ovalbumin, pOVA  =  OVA-peptide, APC  =  T cell-depleted splenocyte, ns  =  not significant. *p<0.05 compared to all other variables tested or as designated by bars, calculated by Mann-Whitney-U test. 2C: ***p<0.001 1way ANOVA with Bonferroni's Multiple Comparison Test Representative experiments of n = 5–12 experiments.</p

IL-4 facilitates airway sensitization by acting on hematopoetic and structural cells.

<p>Chimeric mice, expressing STAT6 in either SC, HC, neither or both were sensitized intranasally with 5 µg OVA in combination with 1 µg IL-4 for three consecutive days and challenged with 25 µg OVA for four days. Animals sensitized with OVA only served as controls. Analyses were performed two days after last challenge. A: BAL cell differentiation of STAT6 chimeric mice. B–E: BAL cell distribution of STAT6 chimeric mice, B  =  eosinophils, C =  lymphocytes, D =  macrophages, E =  neutrophils. F: OVA-specific cytokine expression in lung draining lymph nodes from STAT6 chimeric mice on day 3 of culture. G: OVA-specific IgE in serum of STAT6 chimeric mice. SC  =  structural compartment. HC  =  hematopoetic compartment. LU  =  laboratory units. *p<0.05 by Mann-Whitney-U-test compared to all other variables. Representative experiments of n = 2–8 experiments.</p

Suppression of T cell proliferation by airway epithelial cells is attenuated by IL-4.

<p>Proliferation of OVA-specific DO11.10 T cells was induced by DC-mediated OVA-presentation. Cultures w/o antigen served as controls. Epithelial-mediated suppression was assessed in co-culture with the LA4-cell line, a murine, type II pneumocyte cell line which was pre-treated with the indicated doses of IL-4 (F,G) or pre-treated with 200ng (E) or 10pg of IL-4 (H+I). Harvests were performed on day 4 of co-culture (E+F) or 24 h after IL-4 treatment (H+I). A: mRNA expression of IL-4Rα on untreated LA4 cells, shown are products obtained with different primer concentrations. B: Assessment of IL-4Rα surface expression via immunofluorescence microscopy of untreated LA4 cells, C: Corresponding IL-4Rα surface expression on splenocytes cultured with 1 µg ConA for 3d, D: Corresponding isotype control antibody. E: Pre-treatment with IL-4 (200 ng) reduces epithelial mediated suppression of T cell proliferation on day 4 of co-culture. F: Attenuation of epithelial-dependent suppression of T cell proliferation by IL-4 is dose-dependent. A culture without LA-4 cells (w/o) and IL-4 served as control. G: IL-4 treatment for 2–24 h at the indicated doses does not influence viability of LA4 cells, measured by trypan blue exclusion. H: IL-4-treatment of epithelial cells (10 pg; white) does not change epithelial cytokine secretion (black). I: IL-4 induces GITR-L expression in epithelial cells. TC  =  T cell, DC  =  dendritic cell. *p<0.05 compared to all other variables tested or as designated by bars, calculated by Mann-Whitney-U test. Representative experiments of n = 3–4.</p

Airway epithelial cells interfere with T cell maturation and activation but not by induction of Tregs or modulation of TGF-beta secretion.

<p>Proliferation of OVA-specific DO11.10 T cells was induced by DC-mediated OVA-presentation. Cultures w/o antigen served as controls. Epithelial-mediated suppression was assessed in co-culture with the LA4-cell line, a murine, type II pneumocyte cell line. A: Expression of CD62L vs. CD44 and B: expression of CD25 vs. CD69 was assessed in co-cultures with DCs on day 4 on KJ1-26/CD4+ T cells. C: Secretion of IL-2 in supernatants on day 4 of co-culture with DC as antigen presenting cells. LA4 cells were pretreated 24 h before co-culture with 10 ng/ml IL-4. D: IFN-gamma secretion in supernatants on day 4 of co-cultures with APC or DC as antigen-presenting cells. E: Representative FACS-plot of FoxP3 staining in CD4 T cells from co-cultures of T cells with DCs. F+G: Percentage of FoxP3+/CD4+ T cells in co-cultures with DCs (F) or APC (G) on day 4. H+I: Secretion of TGF-β in co-cultures with DCs (H) or APC (I) on day 4. *p<0.05 compared to all other variables tested or as designated by bars, calculated by Mann-Whitney-U test. 3D: *p<0.05 **p<0.01 ***p<0.001 1-way ANOVA with Bonferronís Multiple Comparison Test. Representative experiments of n = 5–12.</p

Antibiotic susceptibility pattern of the patients’ MRSA isolates (n = 24 patients, n = 26 MRSA isolates).

Arabic numbers indicate patients. For patient 19 and patient 21, two isolates from each two different hospital stays are shown (Roman numbers). Row order is based on the hierarchical clustering of susceptibility patterns using maximum distance and Ward’s linkage.</p

Epidemiological and clinical characteristics of the MRSA patients.

Epidemiological and clinical characteristics of the MRSA patients.</p

Comparison of patients with hospital acquired MRSA (cases, n = 12) and without MRSA (controls, n = 24)—matched case control study.

Comparison of patients with hospital acquired MRSA (cases, n = 12) and without MRSA (controls, n = 24)—matched case control study.</p

Functional lung MRI for regional monitoring of patients with cystic fibrosis

<div><p>Purpose</p><p>To test quantitative functional lung MRI techniques in young adults with cystic fibrosis (CF) compared to healthy volunteers and to monitor immediate treatment effects of a single inhalation of hypertonic saline in comparison to clinical routine pulmonary function tests.</p><p>Materials and methods</p><p>Sixteen clinically stable CF patients and 12 healthy volunteers prospectively underwent two functional lung MRI scans and pulmonary function tests before and 2h after a single treatment of inhaled hypertonic saline or without any treatment. MRI-derived oxygen enhanced T₁ relaxation measurements, fractional ventilation, first-pass perfusion parameters and a morpho-functional CF-MRI score were acquired.</p><p>Results</p><p>Compared to healthy controls functional lung MRI detected and quantified significantly increased ventilation heterogeneity in CF patients. Regional functional lung MRI measures of ventilation and perfusion as well as the CF-MRI score and pulmonary function tests could not detect a significant treatment effect two hours after a single treatment with hypertonic saline in young adults with CF (p>0.05).</p><p>Conclusion</p><p>This study shows the feasibility of functional lung MRI as a non-invasive, radiation-free tool for monitoring patients with CF.</p></div

Receiver operating characteristics analysis.

<p>Receiver operating characteristics analysis.</p

Study protocol.

<p>Pulmonary function testing (spirometry and multiple breath nitrogen washout (MBW)) were performed 60 to 90 minutes prior to the pre treatment scan / 1^st scan. MRI was performed as follows: first, morphological images were assessed followed by phase contrast angiography (PCA) in the ascending aorta. Afterwards T1 mapping breathing room air and again after six minutes of 100% oxygen wash-in time was acquired. Regional Fractional Lung Ventilation MRI (FV) was then acquired under normoxic conditions. Then for assessment of pulmonary parenchymal perfusion, dynamic contrast enhanced (DCE) MRI was carried out followed by a morphological sequence post i.v. contrast. Afterwards inhalation treatment with hypertonic saline (HTS, treatment group) or no treatment (control group) was performed and the PFT (30 minutes after treatment) and MRI (2 h after treatment) were repeated. Healthy volunteers underwent one scan using the same functional lung MRI protocol, except for DCE-MRI and phase-contrast MRI.</p